Abstract
Tissue damage induces rapid recruitment of leukocytes and changes in the transcriptional landscape that influence wound healing. However, the cell-type specific transcriptional changes that influence leukocyte function and tissue repair have not been well characterized. Here, we employed translating ribosome affinity purification (TRAP) and RNA sequencing, TRAP-seq, in larval zebrafish to identify genes differentially expressed in neutrophils, macrophages, and epithelial cells in response to wounding. We identified the complement pathway and c3a.1, homologous to the C3 component of human complement, as significantly increased in neutrophils in response to wounds. c3a.1−/− zebrafish larvae have impaired neutrophil directed migration to tail wounds with an initial lag in recruitment early after wounding. Moreover, c3a.1−/− zebrafish larvae have impaired recruitment to localized bacterial infections and reduced survival that is, at least in part, neutrophil mediated. Together, our findings support the power of TRAP-seq to identify cell type specific changes in gene expression that influence neutrophil behavior in response to tissue damage.
Highlights
Tissue damage induces rapid recruitment of leukocytes and changes in the transcriptional landscape that influence wound healing
We have previously reported the use of translating ribosome affinity purification (TRAP) to detect changes in gene expression in specific cell types resulting from heat shock in zebrafish[9]
We report that cell-specific TRAP-RNAseq of larval zebrafish identifies genes differentially expressed in neutrophils, macrophages, and epithelial cells in response to wounds
Summary
Tissue damage induces rapid recruitment of leukocytes and changes in the transcriptional landscape that influence wound healing. Acute tissue injury is characterized by a rapid influx of both neutrophils and macrophages into the wound microenvironment, followed by inflammatory resolution and wound healing. This initial recruitment of leukocytes to the wound is of critical importance; neutrophils, the most abundant cell type and the first responders to tissue damage, limit infection at the wound s ite[1], while macrophages remove debris that would otherwise impede the repair process[2]. We have previously reported the use of translating ribosome affinity purification (TRAP) to detect changes in gene expression in specific cell types resulting from heat shock in zebrafish[9]. We identified several hundred differentially expressed genes, including upregulation of complement system components
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